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A synthetic peptide corresponding to a sequence near the C-terminal of human Annexin A1, different from the related rat and mouse sequence by two amino acids.
Our Abpromise guarantee covers the use of ab65844 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 - 2 µg/ml. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 39 kDa.|
|IP||Use a concentration of 5 µg/ml.|
Annexin A1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Annexin A1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65844.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 39kDa; Annexin A1, non specific bands - 52kDa and in control (lane 2): We are unsure as to the identity of this extra band.
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